CN110200913B - Preparation method of embedded sulbactam amoxicillin amide compound - Google Patents

Abstract

The invention discloses a preparation method for embedding sulbactam amoxicillinamide complex. First, the protein complex in duck egg white is covalently reacted with glucan in water to obtain a graft, that is, the water phase; The bactam-amoxicillin amide complex is dissolved in soybean oil to obtain an oil phase; and then the water phase and the oil phase are mixed by high-pressure homogenization to prepare a nanoemulsion of the embedded sulbactam-amoxicillin amide complex drug. The nanoemulsion has a stable oil-water interface film and good water solubility, which brings possibilities for the preparation of the above-mentioned complexes; the embedding effect is good, there is no agglomeration or aggregation within 12 months, the particles are uniform and stable, and the particle size is relatively small. Small, can be better absorbed by the body. The raw materials are simple and easy to obtain, safe, low in cost, simple in preparation process, and easy to apply in industrial production, which provides a new idea for the production of water-insoluble pharmaceutical preparations, and can also promote the comprehensive development and utilization of high value-added components in the egg industry, and improve the Economic Value.

Description

A kind of preparation method of embedding sulbactam amoxicillinamide compound

technical field

The invention relates to the technical field of medicine, in particular to a preparation method of an embedded sulbactam-amoxicillinamide compound.

Background technique

Sulbactam is a synthetic and irreversible competitive β-lactamase inhibitor. Its antibacterial activity is weak, slightly stronger than that of clavulanic acid. When used alone, it only has bactericidal effect on Neisseria gonorrhoeae and Acinetobacter. Most β-lactamases produced by blue-positive and gram-negative bacteria have strong inhibitory effects. Amoxicillin has a broad antibacterial spectrum, strong bactericidal power and rapid action. It is widely used in clinical practice because of its oral administration. It can be used to treat typhoid fever, other Salmonella infections, typhoid carriers, urethra, ears, nose, throat, respiratory tract and other diseases. Infection of soft tissue. WHO recommends it as the first-choice β-lactam oral antibiotic, and it has become one of the most eye-catching and fastest-growing antibiotic varieties. However, because the β-lactam structure in the amoxicillin molecule is prone to ring-opening, unstable, prone to degradation, and has poor water solubility, its sodium salt is often used in its injections. Sulbactam also has a certain instability problem. Affected by pH, temperature, oxidation and other factors, its β-lactam structure is also easy to open the ring, resulting in the destruction of the entire molecule, the decrease in content, and the increase in related substances.

In the chemical structure modification of organic drugs, the modification of salt, ester and amide is a kind of common methods. Using two different drug molecules with synergistic functions, after these methods are modified, the structures of the two drugs can be assembled into one molecule, or the pharmacophore of the two can be compatible in one molecule. one of the research interests. In the previous research (CN201210290947.1), the inventors used modification technology to amidate amoxicillin and sulbactam to form sulbactam amoxicillin amide complex, and realized a novel structure of sulbactam The synthesis of moxilinamide complex, its structural formula is as follows:

该酰胺复合物结构上的改造修饰后药物在某种程度上，可以强化药理作用，减小各自相应的毒副作用；或使两者取长补短，发挥各自的药理活性，协同地完成治疗过程；且修饰后更便于拓展临床使用和疾病的治疗与控制。但是，舒巴坦阿莫西林酰胺复合物的水溶解性较低，给制剂生产带来许多不便，限制了其制剂类型特别是水溶液制剂的发展，同时该酰胺复合物仍属β-内酰胺结构，制剂中如何提高其稳定性，也是急需解决的重要问题。

The modified drug on the structure of the amide complex can, to a certain extent, strengthen the pharmacological action and reduce their respective toxic and side effects; Later, it is more convenient to expand clinical use and disease treatment and control. However, the water solubility of the sulbactam-amoxicillin amide complex is low, which brings a lot of inconvenience to the production of preparations, which limits the development of its preparation types, especially the development of aqueous preparations. At the same time, the amide complex is still a β-lactam structure. , how to improve its stability in preparations is also an important problem that needs to be solved urgently.

SUMMARY OF THE INVENTION

In view of the above-mentioned deficiencies of the prior art, the purpose of the present invention is to provide a preparation method for embedding the sulbactam amoxicillin amide compound, which solves the problem of the poor water solubility of the sulbactam amoxicillin amide compound, and it is difficult to prepare a water-soluble compound. Sexual preparations, and the problem of poor structural stability of β-lactam in the complex structure.

In order to solve the above-mentioned technical problems, the present invention adopts the following technical scheme: a preparation method for embedding sulbactam amoxicillin amide compound, comprising the following steps:

1) Dissolve duck egg white in water, then add dextran to obtain a mixed solution, then adjust the pH of the mixed solution to 4-8, stir at room temperature for 1-2 hours to completely dissolve, and place it in a 50°C water bath The reaction is carried out for 1 to 6 hours, cooled to room temperature after the reaction, centrifuged to take the supernatant, and then dialyzed to obtain the graft, that is, the water phase;

2) dissolving the sulbactam amoxicillin amide compound in soybean oil and fully stirring to obtain an oil phase;

3) Mix the water phase obtained in step 1) and the oil phase obtained in step 2), and obtain a nanoemulsion embedding the sulbactam-amoxicillinamide complex by high-pressure homogenization.

Duck egg white is transparent and gelatinous, and mainly contains protein, fat, carbohydrate, calcium, phosphorus, iron, riboflavin, niacin, vitamin A and C and other ingredients. Among them, the proportion of solid matter is 13%, and about 90% of the solid matter is protein, mainly ovalbumin (75%), ovalbumin mostly exists in the form of hydrophilic protein, and most of the hydrophobic amino acids are inside the molecule. , the structure is easily unfolded under the action of high pressure homogenization and calcium, phosphorus, iron and other groups, and interacts with ovomucoid (15%), ovomucoid (7%) and conalbumin (3%) protein complexes, thus endow them with stronger hydrophobic interactions and disulfide bond cross-links between molecules, with the ability to rapidly adsorb to the interface, to stretch and orient rapidly after reaching the interface, and to interact with neighboring molecules after reaching the interface. The interaction forms a film with strong cohesion and viscoelasticity, forming a specific type of protein surfactant; and the synergistic effect of the above protein complex with ovotransferrin and lysozyme in egg white further improves its emulsification effect. The composition of duck egg white is different from other egg whites in terms of protein type, trace elements and other nutrients. These differences in composition may make duck egg white suitable for encapsulating sulbactam-amoxicillinamide compound drugs.

Under the strong shearing, impacting and cavitation generated under high pressure, egg white forms a protein complex with protein-like surfactants, the hydrophilic group of the protein complex is combined with water, and the lipophilic group and oil phase Combined, the complex protein will be bound on the surface of the emulsion droplets to form an oil-water interface film, so as to obtain the immobilization of the oil droplets and complete the efficient loading and entrapment of the sulbactam-amoxicillinamide complex drug, forming countless microscopic or even micron or Nanoscale emulsion. The oil phase and the water phase are uniformly mixed by using the micro-jet generated by the cavitation effect, so that the liquid substance is ultra-fine. At the same time, the glucan grafted on the egg white protein stretches on the surface of the milk droplets due to its hydrophilicity, so that the milk droplets can exist stably in the water for a long time and avoid their precipitation and aggregation.

Preferably, the mass of the duck egg white, water and glucan is 1:2-10:0.05-0.2.

Preferably, the molecular weight of the glucan is 5000-50000.

Preferably, the concentration of the sulbactam-amoxicillinamide compound dissolved in soybean oil in step 2) is 5-100 mg/ml.

Preferably, the mixing volume ratio of the water phase and the oil phase is 1:5-20.

Preferably, the pressure range of the high-pressure homogenization is 300-500 bar, and the high-pressure homogenization time range is 10-30 min.

Preferably, the centrifugal speed is 4000-6000 rpm, and the centrifugal time is 10-20 min.

Preferably, the molecular weight of the dialysis membrane in the dialysis is 10±0.2kDa.

Compared with the prior art, the present invention has the following beneficial effects:

1. The sulbactam amoxicillin amide complex drug-protein complex-dextran-embedded nanoemulsion provided by the present invention is a kind of oil phase containing the sulbactam amoxicillin amide complex drug as the core. , the protein complex in duck egg white is an oil-water interface film, and the dextran grafted on the protein complex is in the outer layer of the nanoemulsion. The nanoemulsion has a stable oil-water interface film, which is stretched on the surface of emulsion droplets due to the hydrophilicity of dextran to avoid the aggregation of emulsion droplets. Well, the stability of the complex is improved, there is no agglomeration or aggregation phenomenon within 12 months, the effective storage period of the sulbactam-amoxicillinamide complex drug is improved, and the drug has a good application prospect.

2. The nanoemulsion particles prepared by the present invention are uniform and stable, and the particle size is small, which can be better absorbed by the human body. When in use, under the action of human digestive enzymes, sulbactam amoxicillinamide compound drug is released along with the degradation of glucan and protein to exert corresponding effects; at the same time, the decomposed glucose and protein peptides also provide nutrition for the human body substance.

3. The present invention uses glucan and duck egg white as raw materials, belongs to conventional food, is non-toxic and easy to degrade, is safe, low in cost, simple in preparation process, short in production cycle, easy to be applied in industrial production, and is also a water-insoluble pharmaceutical preparation The production of egg products provides new ideas, and at the same time can promote the comprehensive development and utilization of high value-added components in the egg industry, and improve the economic value.

Description of drawings

Fig. 1 is a particle size distribution diagram of nanoparticles of sulbactam-amoxicillinamide complex drug-protein complex-dextran nanoemulsion prepared after embedding in Example 1 of the present invention.

Detailed ways

The present invention will be further described in detail below in conjunction with the examples.

Example 1

1) Dissolve duck egg white in 2 times the volume of distilled water in the reaction flask, then add dextran with a molecular weight of 25,000, so that the amount of dextran added is 0.05 times the mass of duck egg white, and then adjust with phosphate buffer solution The pH of the above solution reached 4.0, stirred at room temperature for 2 hours to completely dissolve, and placed it in a water bath at 40 °C for 1 hour. After the reaction was completed, it was cooled to room temperature, centrifuged at 4000 rpm for 15 minutes to remove insoluble matter, and the supernatant was dialyzed at 4 °C for 24 hours ( After dialysis molecular weight 10±0.2kDa), the graft is obtained, which is the water phase;

2) Dissolving the sulbactam-amoxicillin amide complex in soybean oil and stirring sufficiently to make the concentration of the sulbactam-amoxicillin amide complex be 5 mg/ml to obtain an oil phase;

3) Mix the water phase obtained in step 1) and the oil phase obtained in step 2) according to a volume ratio of 5:1, and obtain the embedded sulbactam-amoxicillinamide complex drug-protein complex by high-pressure homogenization A substance-dextran nanoemulsion, wherein the pressure of high-pressure homogenization is 500bar, and the time of high-pressure homogenization is 10min.

Example 2

1) Dissolve duck egg white in 10 times the volume of distilled water in the reaction flask, then add dextran with a molecular weight of 50,000, so that the amount of dextran added is 0.2 times the mass of duck egg white, and then adjust with phosphate buffer solution The pH of the above solution reached 8.0, stirred at room temperature for 2 hours to completely dissolve, and placed it in a 40 °C water bath for 6 hours. After the reaction was completed, it was cooled to room temperature, centrifuged at 6000 rpm for 10 minutes to remove insoluble matter, and the supernatant was dialyzed at 4 °C for 24 hours ( After dialysis molecular weight 10±0.2kDa), the graft is obtained, which is the water phase;

2) dissolving the sulbactam-amoxicillin amide complex in soybean oil and fully stirring, so that the concentration of the sulbactam-amoxicillin amide complex is 100 mg/ml, to obtain an oil phase;

3) Mix the water phase obtained in step 1) and the oil phase obtained in step 2) according to a volume ratio of 20:1, and obtain the embedded sulbactam-amoxicillinamide compound drug-protein compound by high-pressure homogenization A substance-dextran nanoemulsion, wherein the pressure of high-pressure homogenization is 400bar, and the time of high-pressure homogenization is 30min.

Example 3

1) Dissolve duck egg white in 5 times the volume of distilled water in the reaction flask, then add dextran with a molecular weight of 5000, so that the amount of dextran added is 0.1 times the mass of duck egg white, and then adjust with phosphate buffer solution The pH of the above solution reached 5.0, stirred at room temperature for 2 hours to completely dissolve, and placed it in a 40°C water bath for 2 hours of reaction. After the reaction was completed, it was cooled to room temperature, centrifuged at 5000rpm for 20min to remove insoluble matter, and the supernatant was dialyzed at 4°C for 24h ( After dialysis molecular weight 10±0.2kDa), the graft is obtained, which is the water phase;

2) dissolving the sulbactam-amoxicillin amide complex in soybean oil and fully stirring, so that the concentration of the sulbactam-amoxicillin amide complex is 50 mg/ml, to obtain an oil phase;

3) Mix the water phase obtained in step 1) and the oil phase obtained in step 2) according to a volume ratio of 10:1, and obtain the embedded sulbactam-amoxicillinamide complex drug-protein complex by high-pressure homogenization The compound-dextran nanoemulsion, wherein the pressure of high-pressure homogenization is 300bar, and the time of high-pressure homogenization is 20min.

Example 4

1) Dissolve duck egg white in 5 times the volume of distilled water in the reaction flask, then add dextran with a molecular weight of 30,000, so that the amount of dextran added is 0.15 times the mass of duck egg white, and then adjust with phosphate buffer solution The pH of the above solution reached 7.0, stirred at room temperature for 2 hours to completely dissolve, and placed it in a water bath at 40 °C for 1 hour. After the reaction was completed, it was cooled to room temperature, centrifuged at 4000 rpm for 20 minutes to remove insoluble matter, and the supernatant was dialyzed at 4 °C for 24 hours ( After dialysis molecular weight 10±0.2kDa), the graft is obtained, which is the water phase;

2) Dissolving the sulbactam-amoxicillin amide complex in soybean oil and fully stirring, so that the concentration of the sulbactam-amoxicillin amide complex is 20 mg/ml, to obtain an oil phase;

3) Mix the water phase obtained in step 1) and the oil phase obtained in step 2) according to a volume ratio of 10:1, and obtain the embedded sulbactam-amoxicillinamide complex drug-protein complex by high-pressure homogenization A substance-dextran nanoemulsion, wherein the pressure of high-pressure homogenization is 400bar, and the time of high-pressure homogenization is 15min.

Example 5

1) Dissolve duck egg white in 5 times the volume of distilled water in the reaction flask, then add dextran with a molecular weight of 40,000, so that the amount of dextran added is 0.2 times the mass of duck egg white, and then adjust with phosphate buffer solution The pH of the above solution reached 6.0, stirred at room temperature for 2 hours to completely dissolve, and placed it in a 40°C water bath for 1 hour of reaction. After the reaction was completed, it was cooled to room temperature, centrifuged at 4000 rpm for 15 minutes to remove insoluble matter, and the supernatant was dialyzed at 4°C for 24h ( After dialysis molecular weight 10±0.2kDa), the graft is obtained, which is the water phase;

2) dissolving the sulbactam-amoxicillin amide complex in soybean oil and fully stirring, so that the concentration of the sulbactam-amoxicillin amide complex is 50 mg/ml, to obtain an oil phase;

3) Mix the water phase obtained in step 1) and the oil phase obtained in step 2) according to a volume ratio of 15:1, and obtain the embedded sulbactam-amoxicillinamide complex drug-protein complex by high-pressure homogenization A substance-dextran nanoemulsion, wherein the pressure of high-pressure homogenization is 500bar, and the time of high-pressure homogenization is 10min.

Example 6

1) Dissolve duck egg white in 10 times the volume of distilled water in the reaction flask, then add dextran with a molecular weight of 6000, so that the amount of dextran added is 0.05 times the mass of duck egg white, and then adjust with phosphate buffer solution The pH of the above solution reached 4.0, stirred at room temperature for 2 hours to completely dissolve, and placed it in a water bath at 40 °C for 2.5 hours. After the reaction was completed, it was cooled to room temperature, centrifuged at 6000 rpm for 10 minutes to remove insoluble matter, and the supernatant was dialyzed at 4 °C for 24 hours. (The molecular weight of dialysis is 10±0.2kDa) to obtain the graft, which is the water phase;

2) dissolving the sulbactam-amoxicillin amide complex in soybean oil and fully stirring, so that the concentration of the sulbactam-amoxicillin amide complex is 50 mg/ml, to obtain an oil phase;

3) Mix the water phase obtained in step 1) and the oil phase obtained in step 2) according to a volume ratio of 10:1, and obtain the embedded sulbactam-amoxicillinamide complex drug-protein complex by high-pressure homogenization The compound-dextran nanoemulsion, wherein the pressure of high-pressure homogenization is 300bar, and the time of high-pressure homogenization is 20min.

Example 7

1) Dissolve duck egg white in 2 times the volume of distilled water in the reaction flask, then add dextran with a molecular weight of 10,000, so that the amount of dextran added is 0.2 times the mass of duck egg white, and then adjust with phosphate buffer solution The pH of the above solution reached 4.0, stirred at room temperature for 2 hours until completely dissolved, and placed it in a 40 °C water bath for 5 hours. After the reaction was completed, it was cooled to room temperature, centrifuged at 6000 rpm for 20 minutes to remove insoluble matter, and the supernatant was dialyzed at 4 °C for 24 hours ( After dialysis molecular weight 10±0.2kDa), the graft is obtained, which is the water phase;

2) dissolving the sulbactam-amoxicillin amide complex in soybean oil and fully stirring, so that the concentration of the sulbactam-amoxicillin amide complex is 50 mg/ml, to obtain an oil phase;

3) Mix the water phase obtained in step 1) and the oil phase obtained in step 2) according to a volume ratio of 10:1, and obtain the embedded sulbactam-amoxicillinamide complex drug-protein complex by high-pressure homogenization The compound-dextran nanoemulsion, wherein the pressure of high-pressure homogenization is 300bar, and the time of high-pressure homogenization is 20min.

Example 8

1) Dissolve duck egg white in 5 times the volume of distilled water in the reaction flask, then add dextran with a molecular weight of 25,000, so that the amount of dextran added is 0.2 times the mass of duck egg white, and then adjust with phosphate buffer solution The pH of the above solution reached 5.0, stirred at room temperature for 2 hours until completely dissolved, and placed it in a 40 °C water bath for 5 hours. After the reaction was completed, it was cooled to room temperature, centrifuged at 4000 rpm for 20 minutes to remove insoluble matter, and the supernatant was dialyzed at 4 °C for 24 hours ( After dialysis molecular weight 10±0.2kDa), the graft is obtained, which is the water phase;

2) dissolving the sulbactam-amoxicillin amide complex in soybean oil and fully stirring, so that the concentration of the sulbactam-amoxicillin amide complex is 50 mg/ml, to obtain an oil phase;

3) Mix the water phase obtained in step 1) and the oil phase obtained in step 2) according to a volume ratio of 5:1, and obtain the embedded sulbactam-amoxicillinamide complex drug-protein complex by high-pressure homogenization The substance-dextran nanoemulsion, wherein the pressure of high-pressure homogenization is 300bar, and the time of high-pressure homogenization is 30min.

Example 9

1) Dissolve duck egg white in 2 times the volume of distilled water in the reaction flask, then add dextran with a molecular weight of 10,000, so that the amount of dextran added is 0.2 times the mass of duck egg white, and then adjust with phosphate buffer solution The pH of the above solution reached 7.0, stirred at room temperature for 2 hours to completely dissolve, and placed it in a 40 °C water bath for 5 hours. After the reaction was completed, it was cooled to room temperature, centrifuged at 4000 rpm for 15 minutes to remove insoluble matter, and the supernatant was dialyzed at 4 °C for 24 hours ( After dialysis molecular weight 10±0.2kDa), the graft is obtained, which is the water phase;

2) dissolving the sulbactam-amoxicillin amide complex in soybean oil and fully stirring, so that the concentration of the sulbactam-amoxicillin amide complex is 50 mg/ml, to obtain an oil phase;

3) Mix the water phase obtained in step 1) and the oil phase obtained in step 2) according to a volume ratio of 10:1, and obtain the embedded sulbactam-amoxicillinamide complex drug-protein complex by high-pressure homogenization A substance-dextran nanoemulsion, wherein the pressure of high-pressure homogenization is 500bar, and the time of high-pressure homogenization is 20min.

The sulbactam-amoxicillinamide complex drug-protein complex-dextran nano-aqueous emulsion prepared in Example 1 was tested for the particle size of the emulsion by a laser particle size analyzer, and the results are shown in Figure 1. As can be seen from the figure, the particle size of the sulbactam-amoxicillinamide complex drug-protein complex-dextran nano-water emulsion prepared by the present invention is about 237.7±9.31nm, and the emulsion can be passed through a 450nm filter membrane to achieve purpose of sterilization.

The particle size and drug loading of the sulbactam-amoxicillinamide complex drug-protein complex-dextran nano-emulsion prepared in Examples 1-9 were measured at different time periods, and the results are shown in Table 1. .

Table 1

As can be seen from Table 1, in different time periods, the nanoemulsion prepared by the present invention has no significant change in the drug loading of the sulbactam-amoxicillinamide compound drug, indicating that it has a good embedding effect. There is no agglomeration or aggregation within a month, indicating that it has good stability; the particles are uniform and stable, and there is no agglomeration in the aqueous solution, indicating that the prepared nanoemulsion has good water solubility, and the particle size is small and easy to absorb.

The above descriptions are only preferred embodiments of the present invention, and are not intended to limit the present invention. Any modifications, equivalent replacements and improvements made within the spirit and principles of the present invention shall be included in the scope of the present invention. within the scope of protection.

Claims (7)

1. A preparation method of an embedded sulbactam amoxicillin amide compound is characterized by comprising the following steps:

1) dissolving duck egg white in water, adding glucan to obtain a mixed solution, adjusting the pH of the mixed solution to 4-8, stirring at room temperature for 1-2 h until the mixed solution is completely dissolved, placing the mixed solution in a water bath at 50 ℃ for reaction for 1-6 h, cooling to room temperature after the reaction is finished, centrifuging to obtain a supernatant, and dialyzing to obtain a water phase; the duck egg white, water and glucan are 1: 2-10: 0.05 to 0.2;

2) dissolving the sulbactam amoxicillin amide compound in the soybean oil, and fully stirring to obtain an oil phase;

3) mixing the water phase obtained in the step 1) and the oil phase obtained in the step 2), and carrying out high-pressure homogenization to obtain the sulbactam amoxicillin amide compound embedded nano emulsion.

2. The method for preparing the embedded sulbactam amoxicillin amide compound as claimed in claim 1, wherein the molecular weight of the glucan is 5000-50000.

3. The method for preparing the embedded sulbactam amoxicillin amide compound as claimed in claim 1, wherein the concentration of the sulbactam amoxicillin amide compound in the step 2) dissolved in the soybean oil is 5-100 mg/ml.

4. The preparation method of the embedded sulbactam amoxicillin amide compound as claimed in claim 1, wherein the mixing volume ratio of the water phase and the oil phase is 1: 5-20.

5. The method for preparing the embedded sulbactam amoxicillin amide compound as claimed in claim 1, wherein the high pressure homogenization pressure is 300-500 bar, and the high pressure homogenization time is 10-30 min.

6. The method for preparing the embedded sulbactam amoxicillin amide compound as claimed in claim 1, wherein the centrifugation speed is 4000-6000 rpm, and the centrifugation time is 10-20 min.

7. The method for preparing the embedded sulbactam amoxicillin amide complex as claimed in claim 1, wherein the dialyzed molecular weight is 10 ± 0.2 kDa.

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